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Method for fermentation production of lipstatin and culture medium components thereof

A fermentation medium and a fermentation culture technology are applied in the field of methods and medium components, fermentation production of liprastatin, which can solve the problems of low fermentation level and instability, and achieve the purpose of reducing fermentation cost, improving stability and improving The effect of fermentation titer

Active Publication Date: 2013-06-12
鲁南新时代生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problem of low and unstable fermentation level in the prior art, the present invention provides a new medium for fermentation and production of riprestatin, a method for controlling the fermentation process and components of the fermentation medium

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Incline cultivation:

[0019] Prepare the medium according to the following ratio: glucose 2g / L, malt extract 8g / L, yeast extract 10g / L, agar 15g / L, pH 7.1. Prepare 300 ml of medium, heat the agar to melt, stir evenly, put it into 10 large test tubes, sterilize it, and make it into a slope for later use. The lyophilized Streptomyces toxin strain was dissolved with 5 milliliters of sterile water, and 4 milliliters were evenly spread on 10 inclined surfaces at a temperature of 28° C. and a humidity of 40 to 60%, and cultivated for 10 days to produce a large number of spores. Add sterile water to make a spore suspension.

[0020] Shake flask seed solution preparation:

[0021] The medium was prepared according to the following proportions: glycerol 10g / L, malt extract 10g / L, yeast extract 5g / L, defatted soybean powder 20g / L, pH 6.0. Prepare 200 ml of culture medium, put it into 4 bottles of 500 mL Erlenmeyer flasks (each bottle contains 50 mL), sterilize and set aside. ...

Embodiment 2

[0027] Incline cultivation:

[0028] The medium was prepared according to the following proportions: glucose 8g / L, malt extract 4g / L, yeast extract 4g / L, agar 14g / L, pH 7.4. Prepare 300 ml of medium, heat the agar to melt, stir evenly, put it into 10 large test tubes, sterilize it, and make it into a slope for later use. The lyophilized Streptomyces toxin strain was dissolved with 5 milliliters of sterile water, and 4 milliliters were evenly spread on 10 inclined surfaces at a temperature of 28° C. and a humidity of 40 to 60%, and cultivated for 8 days to produce a large number of spores. Add sterile water to make a spore suspension.

[0029] Shake flask seed solution preparation:

[0030] The medium was prepared according to the following proportions: glycerol 10g / L, malt extract 10g / L, yeast extract 5g / L, defatted soybean powder 20g / L, pH 6.0. Prepare 200 ml of culture medium, put it into 4 bottles of 500 mL Erlenmeyer flasks (each bottle contains 50 mL), sterilize and se...

Embodiment 3

[0036] Incline cultivation:

[0037] The medium was prepared according to the following proportions: glucose 8g / L, malt extract 2g / L, yeast extract 4g / L, agar 12g / L, pH 7.2. Prepare 300 ml of medium, heat the agar to melt, stir evenly, put it into 10 large test tubes, sterilize it, and make it into a slope for later use. The lyophilized Streptomyces toxin strain was dissolved with 5 milliliters of sterile water, and 4 milliliters were evenly spread on 10 inclined surfaces at a temperature of 28° C. and a humidity of 40 to 60%, and cultivated for 10 days to produce a large number of spores. Add sterile water to make a spore suspension.

[0038] Shake flask seed solution preparation:

[0039] The medium was prepared according to the following proportions: glycerol 10g / L, malt extract 10g / L, yeast extract 5g / L, defatted soybean powder 20g / L, pH 6.0. Prepare 200 ml of culture medium, put it into 4 bottles of 500 mL Erlenmeyer flasks (each bottle contains 50 mL), sterilize and s...

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PUM

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Abstract

The invention provides a method for fermentation production of lipstatin, belonging to the field of fermentation production of microorganisms. The method takes streptomyces toxytricini as an initial bacterium and mainly comprises the steps of preparation of a slope strain, preparation of a seed solution, control of a fermentation process and the like. In the invention, by taking soybean meal, bean oil and glycerol as main nitrogen sources and carbon sources of a fermentation culture medium and regulating and controlling the fermentation process and the components of the culture medium, the production cost of the lipstatin is greatly lowered, and the fermentation titer is increased, thus the defects of high production cost and low fermentation titer in the prior art are overcome.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a method for fermenting and producing riprestatin and its culture medium components. Background technique [0002] The liprestatin produced by the fermentation method involved in the present invention is an intermediate of the novel weight-loss drug orlistat. Orlistat is currently the only weight-loss drug in the world. It is a long-acting and potent specific gastrointestinal lipase inhibitor. The covalent bond inactivates the enzyme to play a therapeutic role. The inactivated enzyme cannot hydrolyze the fat in food, mainly triglyceride, into absorbable free fatty acid and monoacylglycerol. Undigested triglycerides cannot be absorbed by the body, thereby reducing caloric intake and controlling weight. [0003] There are two methods for the production of orlistat, fermentation synthesis and total synthesis. The production cost of the total synthesis method is relatively hi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P17/02C12R1/465
Inventor 赵志全刘茂田
Owner 鲁南新时代生物技术有限公司
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